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Background  The objective of this work was to engineer  Deinococcus radiodurans  R1 as a microbial cell factory for the production of pinene, a monoterpene molecule prominently used for the production of fragrances, pharmaceutical products, and jet engine biofuels. Our objective was to produce pinene from glycerol, an abundant by-product of various industries.    Results  To enable pinene production in  D. radiodurans , we expressed the pinene synthase from  Abies grandis , the geranyl pyrophosphate (GPP) synthase from  Escherichia coli , and overexpressed the native 1-deoxy- d -xylulose 5-phosphate synthase. Further, we disrupted the deinoxanthin pathway competing for the substrate GPP by either inactivating the gene  dr0862 , encoding phytoene synthase, or substituting the native GPP synthase with that of  E. coli . These manipulations resulted in a  D. radiodurans  strain capable of producing 3.2 ± 0.2 mg/L pinene in a minimal medium supplemented with glycerol, with a yield of 0.13 ± 0.04 mg/g glycerol in shake flask cultures. Additionally, our results indicated a higher tolerance of  D. radiodurans  towards pinene as compared to  E. coli .    Conclusions  In this study, we successfully engineered the extremophile bacterium  D. radiodurans  to produce pinene. This is the first study demonstrating the use of  D. radiodurans  as a cell factory for the production of terpenoid molecules. Besides, its high resistance to pinene makes  D. radiodurans  a suitable host for further engineering efforts to increase pinene titer as well as a candidate for the production of the other terpenoid molecules.    Supplementary Information  The online version contains supplementary material available at 10.1186/s12934-021-01674-4.

Metabolic engineering of Deinococcus radiodurans for pinene production from glycerol